TET1-mediated 5-hydroxymethylcytosine modification & airway hyperresponsiveness

TET1介导的5-羟甲基胞嘧啶修饰

基本信息

批准号：
9493470
负责人：
Wan-yee Tang
金额：
$ 34.43万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9493470
关键词：
Aberrant DNA Methylation Acute Address Adult Affect Allergens Allergic Antioxidants Asthma Automobile Driving B-Lymphocytes Biological Assay Biology Bronchodilation Carbon Carbon Black Cell Proliferation Cell physiology Cells Chronic Chronic Disease Collagen Complex Cytosine DNA DNA Methylation DNA Sequence Data Deposition Development Diesel Exhaust Dioxygenases Disease Environmental Risk Factor Epigenetic Process Etiology Excision Exposure to Gene Expression Genes Genetic Genetic Transcription Heritability Human Immunologics In Vitro Lead Link Lung Mediating Methionine Mitotic Modification Mus Muscle function NADH Organ Oxidation-Reduction Oxidative Stress Oxides Particulate Matter Pathogenesis Phenotype Physiological Physiology Play Prevalence Process Production Protein translocation Proteins Pulmonology Pyroglyphidae Regulation Research Personnel Respiratory physiology Risk Role Signaling Molecule Smooth Muscle Myocytes Structure of parenchyma of lung Sulforaphane T-Lymphocyte Techniques Tetanus Helper Peptide Time Tissues Translating Untranslated RNA Up-Regulation Validation airway hyperresponsiveness airway inflammation airway remodeling alpha ketoglutarate asthmatic asthmatic airway base cell type cigarette smoke epigenetic regulation experimental study gene environment interaction histone modification in vivo innovation insight methyl group methylation pattern mouse model multidisciplinary next generation sequencing novel novel therapeutic intervention oxidation postnatal prenatal public health relevance respiratory smooth muscle

项目摘要

DESCRIPTION (provided by applicant): Asthma is a complex disease characterized by airway hyperresponsiveness (AHR) and airway inflammation, which current afflicts over 300 million people worldwide. Although genetic factors unquestionably play a role in asthma, the rapid rise in asthma prevalence suggests that environmental factors likely play an equally important role. To date, epigenetic regulation is suggested to mediate, at least partly, the complex gene-by- environment interactions that can lead to asthma. Most studies to date have focused on elucidation of DNA methylation patterns in easily accessible cells including T cells and B cells to explain the immunological mechanisms driving allergen sensitization in asthmatics. Few studies have examined the epigenetic consequences of environmental triggers on lung tissues linked to chronic alterations in lung functions. We have utilized next generation sequencing techniques and proper validation assays to identify a novel set of genes which is highly enriched for Tgfb2 signaling molecules and associated with exposure to the common allergen, house dust mite (HDM), and the development of AHR through epigenetic modulation of the airway smooth muscle (ASM) cell phenotypes. In addition, we identified a novel role of the specific 5-mC dioxygenase, TET1, in the regulation of ASM function in vitro and in the development of allergen-driven AHR in vivo. Preliminarily, we demonstrated Tet1 deficiency reduced acute HDM-driven AHR in mice. Strikingly, we were able to translate the epigenetic changes in human asthmatic ASM cells; showing Tet1-mediated hydroxymethylation may influence ASM cell proliferation and contraction. Our study represents the first demonstration that Tet1 protein is regulated in the context of allergen exposure, although the mechanisms regulating Tet1 induction by allergens are unknown. Our preliminary data supported that oxidative stress generated by HDM exposure activates Tet1 and its mediated upregulation of ASM genes. Based on these novel findings, we propose the central hypothesis "HDM activates Tet1-mediated hydroxymethylation in ASM cells through regulation of redox cycling. This epigenetic reprogramming persists in ASM over long time spans, thus likely contributes to the increased ASM cell proliferation, stiffness and contractility seen in human asthma". To address these novel hypotheses, we have assembled a multi-disciplinary team of investigators with a breadth of expertise spanning the fields of epigenetics, ASM biology, pulmonology and pathobiology. First, we will determine the role of Tet1- mediated DNA hydroxymethylation in the development of the AHR phenotype in a chronic HDM-induced AHR mouse model. Second, we will investigate how HDM alters the Tet1 activity, through redox cycling of NADH in both in vitro and in vivo mouse model. Lastly, we will confirm if TET1-mediated hydroxymethylation regulates human ASM cell phenotypes. The results of the proposed experiments should provide novel perspectives on the ASM and its contributions to the excessive airway narrowing in asthma. These insights may fuel the development of new therapeutic approaches for the treatment of this debilitating disease.

描述（由适用提供）：哮喘是一种复杂的疾病，其特征是气道高反应性（AHR）和气道感染，目前影响全球超过3亿人。尽管遗传因素无疑在哮喘中起作用，但哮喘患病率的迅速升高表明环境因素可能起着同样重要的作用。迄今为止，建议表观遗传调节至少部分地介导可能导致哮喘的复杂基因相互作用。迄今为止，大多数研究都集中在易于访问的细胞中的DNA甲基化模式（包括T细胞和B细胞）解释在哮喘患者中驱动过敏原传感器的免疫机制。很少有研究检查了环境触发因素对与肺功能长期改变有关的肺组织的表观遗传后果。我们已经利用了下一代测序技术和适当的验证方法来识别一组新型的基因，该基因高度丰富了TGFB2信号分子，并且与暴露于公共过敏原，房屋粉尘Mitte（HDM）以及通过空中平滑肌（ASM）平滑肌（ASM）平滑肌肉的表观遗传学的aHR开发有关。此外，我们确定了特定5-MC二氧酶TET1在体外和过敏原驱动的AHR在体内的发展中的新作用。初步的，我们证明了TET1缺乏症减少了小鼠急性HDM驱动的AHR。令人惊讶的是，我们能够翻译人类哮喘ASM细胞中的表观遗传变化。显示TET1介导的羟甲基可能会影响ASM细胞增殖和收缩。我们的研究代表了第一次证明TET1蛋白在过敏原暴露的情况下受到调节，尽管该机理调节了过敏原诱导TET1诱导。我们的初步数据支持HDM暴露产生的氧化应激激活TET1及其介导的ASM基因上调。基于这些新发现，我们提出了中心假设“ HDM通过调节氧化还原循环的调节来激活ASM细胞中TET1介导的羟甲基化。这种表观遗传学重编程在长期跨度中仍然存在于ASM中，因此可能有助于增加的ASM细胞增殖，僵硬，僵硬，僵硬，僵硬，僵硬，僵硬，僵硬，僵硬，僵硬，并在人类Asthma中看到。为了解决这些新颖的假设，我们组建了一个多学科的研究人员团队，具有跨越表观遗传学，ASM生物学，肺病学和病理生物学领域的广泛专业知识。首先，我们将确定慢性HDM诱导的AHR小鼠模型中TET1介导的DNA羟基甲基化在AHR表型中的作用。其次，我们将通过体外和体内小鼠模型中NADH的氧化还原循环来研究HDM如何改变TET1活性。最后，我们将确认TET1介导的羟甲基化是否调节人ASM细胞表型。拟议的实验的结果应提供有关ASM及其对哮喘中过度气道的贡献的新观点。这些见解可能会开发出新的治疗方法来治疗这种使人衰弱的疾病。